This is the same notebook as 100_species, but we look at the wadjemup/rottnest queryset. This should be slightly harder. # Setup

knitr::opts_chunk$set(warning = FALSE)

source(here::here('code', 'helpers.R'))
library(tidyverse)
library(forcats)
library(cowplot)
library(ggupset)
library(RColorBrewer)
library(patchwork)
library(vegan)
library(pheatmap)
library(broom)
data <- targets::tar_read(merged_all_results)
# rename BLAST to BLAST97 to differentiate from BLAST100 (percentage identity in both cases)
data <- data |> mutate(Type = str_replace(Type, '^BLAST$', 'BLAST97'))
truth <- targets::tar_read(truth_set_data)
table(data$Type)
## 
##    BLAST100     BLAST97   CustomNBC   DADA2Spec    DADA2Tax  Kraken_0.0 
##       13912       21093      155700      157104      157104      104880 
## Kraken_0.05  Kraken_0.1    Metabuli MMSeqs2_100  MMSeqs2_97      Mothur 
##      104880      104880       73217      109368      109368      152892 
##      Qiime2     VSEARCH 
##       23275      157104

Let’s remove the >0.2 Kraken runs, those are too strict

data <- data |> filter(!Type %in% c('Kraken_0.2', 'Kraken_0.3', 'Kraken_0.4', 'Kraken_0.5', 'Kraken_0.6', 'Kraken_0.7', 'Kraken_0.8', 'Kraken_0.9'))

Made a mistake- Metabuli’s and TNT’s databases is misspelled

data <- data |> mutate(Subject = str_replace_all(Subject, pattern = '_ref.fasta', replacement=''))
data <- data |> mutate(Subject = str_replace_all(Subject, pattern = 'final.csv', replacement = 'final.fasta'))
table(data$Query)
## 
##                                                                                KWest_16S_PooledTrue.fa 
##                                                                                                 366180 
##   make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_12S_Lulu_RESULTS_dada2_asv.fa 
##                                                                                                 100033 
## make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_12S_noLulu_RESULTS_dada2_asv.fa 
##                                                                                                  99769 
##   make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_16S_Lulu_RESULTS_dada2_asv.fa 
##                                                                                                  97434 
## make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_16S_noLulu_RESULTS_dada2_asv.fa 
##                                                                                                  97254 
##        make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_CO1_RESULTS_dada2_asv.fa 
##                                                                                                 101106 
##                               make_12s_16s_simulated_reads_6-fakeGenes_GreenGenes_RESULTS_dada2_asv.fa 
##                                                                                                  88422 
##                                   make_12s_16s_simulated_reads_6-fakeGenes_Random_RESULTS_dada2_asv.fa 
##                                                                                                  89094 
##                     make_12s_16s_simulated_reads_7-Lutjanids_Mock_runEDNAFlow_12S_RESULTS_dada2_asv.fa 
##                                                                                                  24355 
##                     make_12s_16s_simulated_reads_7-Lutjanids_Mock_runEDNAFlow_16S_RESULTS_dada2_asv.fa 
##                                                                                                  26746 
##                     make_12s_16s_simulated_reads_7-Lutjanids_Mock_runEDNAFLOW_CO1_RESULTS_dada2_asv.fa 
##                                                                                                  26505 
##                           make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_12S_RESULTS_dada2_asv.fa 
##                                                                                                 102811 
##                           make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_16S_RESULTS_dada2_asv.fa 
##                                                                                                 109807 
##                           make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_CO1_RESULTS_dada2_asv.fa 
##                                                                                                 115261

Wadjemup species

Check 12S results

table(data$Subject)
## 
##                                                                                            12s_v010_final.fasta 
##                                                                                                           16590 
##                                            12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_1.fasta 
##                                                                                                           16175 
##                                           12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_10.fasta 
##                                                                                                           15914 
##                                            12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_2.fasta 
##                                                                                                           15918 
##                                            12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_3.fasta 
##                                                                                                           16012 
##                                            12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_4.fasta 
##                                                                                                           16089 
##                                            12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_5.fasta 
##                                                                                                           16203 
##                                            12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_6.fasta 
##                                                                                                           15855 
##                                            12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_7.fasta 
##                                                                                                           15966 
##                                            12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_8.fasta 
##                                                                                                           16261 
##                                            12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_9.fasta 
##                                                                                                           16047 
##                                          12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_1.fasta 
##                                                                                                           16366 
##                                         12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_10.fasta 
##                                                                                                           16276 
##                                          12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_2.fasta 
##                                                                                                           16170 
##                                          12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_3.fasta 
##                                                                                                           16336 
##                                          12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_4.fasta 
##                                                                                                           16075 
##                                          12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_5.fasta 
##                                                                                                           16297 
##                                          12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_6.fasta 
##                                                                                                           15985 
##                                          12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_7.fasta 
##                                                                                                           16148 
##                                          12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_8.fasta 
##                                                                                                           16105 
##                                          12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_9.fasta 
##                                                                                                           16282 
##                                           12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_1.fasta 
##                                                                                                           15716 
##                                          12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_10.fasta 
##                                                                                                           15455 
##                                           12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_2.fasta 
##                                                                                                           15781 
##                                           12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_3.fasta 
##                                                                                                           15799 
##                                           12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_4.fasta 
##                                                                                                           15581 
##                                           12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_5.fasta 
##                                                                                                           15704 
##                                           12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_6.fasta 
##                                                                                                           15553 
##                                           12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_7.fasta 
##                                                                                                           15589 
##                                           12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_8.fasta 
##                                                                                                           15583 
##                                           12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_9.fasta 
##                                                                                                           15626 
##                                                                        12s_v010_finalforTaxonomy.filtered.fasta 
##                                                                                                             240 
##   12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_10forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_1forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_2forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_3forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_4forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_5forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_6forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_7forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_8forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_9forTaxonomy.filtered.fasta 
##                                                                                                             240 
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_10forTaxonomy.filtered.fasta 
##                                                                                                             240 
##  12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_1forTaxonomy.filtered.fasta 
##                                                                                                             240 
##  12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_2forTaxonomy.filtered.fasta 
##                                                                                                             240 
##  12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_3forTaxonomy.filtered.fasta 
##                                                                                                             240 
##  12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_4forTaxonomy.filtered.fasta 
##                                                                                                             240 
##  12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_5forTaxonomy.filtered.fasta 
##                                                                                                             240 
##  12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_6forTaxonomy.filtered.fasta 
##                                                                                                             240 
##  12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_7forTaxonomy.filtered.fasta 
##                                                                                                             240 
##  12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_8forTaxonomy.filtered.fasta 
##                                                                                                             240 
##  12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_9forTaxonomy.filtered.fasta 
##                                                                                                             240 
##  12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_10forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_1forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_2forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_3forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_4forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_5forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_6forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_7forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_8forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_9forTaxonomy.filtered.fasta 
##                                                                                                             240 
##                                                                                            12S_v10_HmmCut.fasta 
##                                                                                                            8714 
##                                                                                             16S_v04_final.fasta 
##                                                                                                           17674 
##                                             16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_1.fasta 
##                                                                                                           16374 
##                                            16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_10.fasta 
##                                                                                                           16360 
##                                             16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_2.fasta 
##                                                                                                           16735 
##                                             16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_3.fasta 
##                                                                                                           16567 
##                                             16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_4.fasta 
##                                                                                                           16776 
##                                             16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_5.fasta 
##                                                                                                           16689 
##                                             16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_6.fasta 
##                                                                                                           16330 
##                                             16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_7.fasta 
##                                                                                                           16841 
##                                             16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_8.fasta 
##                                                                                                           16165 
##                                             16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_9.fasta 
##                                                                                                           16285 
##                                           16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_1.fasta 
##                                                                                                           17050 
##                                          16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_10.fasta 
##                                                                                                           16867 
##                                           16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_2.fasta 
##                                                                                                           17165 
##                                           16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_3.fasta 
##                                                                                                           16830 
##                                           16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_4.fasta 
##                                                                                                           17045 
##                                           16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_5.fasta 
##                                                                                                           17369 
##                                           16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_6.fasta 
##                                                                                                           16953 
##                                           16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_7.fasta 
##                                                                                                           17104 
##                                           16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_8.fasta 
##                                                                                                           17262 
##                                           16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_9.fasta 
##                                                                                                           17159 
##                                            16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_1.fasta 
##                                                                                                           16172 
##                                           16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_10.fasta 
##                                                                                                           16327 
##                                            16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_2.fasta 
##                                                                                                           16294 
##                                            16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_3.fasta 
##                                                                                                           16191 
##                                            16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_4.fasta 
##                                                                                                           16435 
##                                            16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_5.fasta 
##                                                                                                           16174 
##                                            16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_6.fasta 
##                                                                                                           15983 
##                                            16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_7.fasta 
##                                                                                                           16145 
##                                            16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_8.fasta 
##                                                                                                           16010 
##                                            16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_9.fasta 
##                                                                                                           16118 
##                                                                         16S_v04_finalforTaxonomy.filtered.fasta 
##                                                                                                             240 
##    16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_10forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_1forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_2forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_3forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_4forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_5forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_6forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_7forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_8forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_9forTaxonomy.filtered.fasta 
##                                                                                                             240 
##  16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_10forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_1forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_2forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_3forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_4forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_5forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_6forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_7forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_8forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_9forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_10forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_1forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_2forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_3forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_4forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_5forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_6forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_7forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_8forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_9forTaxonomy.filtered.fasta 
##                                                                                                             240 
##                                                                                            16S_v04_HmmCut.fasta 
##                                                                                                            9590 
##                                                                                             c01_v03_final.fasta 
##                                                                                                           12609 
##                                             c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_1.fasta 
##                                                                                                           12286 
##                                            c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_10.fasta 
##                                                                                                           12907 
##                                             c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_2.fasta 
##                                                                                                           12333 
##                                             c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_3.fasta 
##                                                                                                           12919 
##                                             c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_4.fasta 
##                                                                                                           12107 
##                                             c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_5.fasta 
##                                                                                                           12272 
##                                             c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_6.fasta 
##                                                                                                           12223 
##                                             c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_7.fasta 
##                                                                                                           12673 
##                                             c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_8.fasta 
##                                                                                                           12229 
##                                             c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_9.fasta 
##                                                                                                           12925 
##                                           c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_1.fasta 
##                                                                                                           12455 
##                                          c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_10.fasta 
##                                                                                                           12383 
##                                           c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_2.fasta 
##                                                                                                           12356 
##                                           c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_3.fasta 
##                                                                                                           12334 
##                                           c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_4.fasta 
##                                                                                                           12420 
##                                           c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_5.fasta 
##                                                                                                           12446 
##                                           c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_6.fasta 
##                                                                                                           12329 
##                                           c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_7.fasta 
##                                                                                                           12419 
##                                           c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_8.fasta 
##                                                                                                           12572 
##                                           c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_9.fasta 
##                                                                                                           12499 
##                                            c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_1.fasta 
##                                                                                                           12989 
##                                           c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_10.fasta 
##                                                                                                           12948 
##                                            c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_2.fasta 
##                                                                                                           12809 
##                                            c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_3.fasta 
##                                                                                                           12053 
##                                            c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_4.fasta 
##                                                                                                           12010 
##                                            c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_5.fasta 
##                                                                                                           12513 
##                                            c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_6.fasta 
##                                                                                                           12803 
##                                            c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_7.fasta 
##                                                                                                           12641 
##                                            c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_8.fasta 
##                                                                                                           11991 
##                                            c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_9.fasta 
##                                                                                                           12002 
##                                                                         c01_v03_finalforTaxonomy.filtered.fasta 
##                                                                                                             240 
##    c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_10forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_1forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_2forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_3forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_4forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_5forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_6forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_7forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_8forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_9forTaxonomy.filtered.fasta 
##                                                                                                             240 
##  c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_10forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_1forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_2forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_3forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_4forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_5forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_6forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_7forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_8forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_9forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_10forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_1forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_2forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_3forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_4forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_5forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_6forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_7forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_8forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_9forTaxonomy.filtered.fasta 
##                                                                                                             240 
##                                                                                            c01_v03_HmmCut.fasta 
##                                                                                                            6792
table(data$Subject)
## 
##                                                                                            12s_v010_final.fasta 
##                                                                                                           16590 
##                                            12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_1.fasta 
##                                                                                                           16175 
##                                           12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_10.fasta 
##                                                                                                           15914 
##                                            12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_2.fasta 
##                                                                                                           15918 
##                                            12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_3.fasta 
##                                                                                                           16012 
##                                            12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_4.fasta 
##                                                                                                           16089 
##                                            12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_5.fasta 
##                                                                                                           16203 
##                                            12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_6.fasta 
##                                                                                                           15855 
##                                            12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_7.fasta 
##                                                                                                           15966 
##                                            12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_8.fasta 
##                                                                                                           16261 
##                                            12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_9.fasta 
##                                                                                                           16047 
##                                          12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_1.fasta 
##                                                                                                           16366 
##                                         12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_10.fasta 
##                                                                                                           16276 
##                                          12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_2.fasta 
##                                                                                                           16170 
##                                          12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_3.fasta 
##                                                                                                           16336 
##                                          12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_4.fasta 
##                                                                                                           16075 
##                                          12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_5.fasta 
##                                                                                                           16297 
##                                          12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_6.fasta 
##                                                                                                           15985 
##                                          12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_7.fasta 
##                                                                                                           16148 
##                                          12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_8.fasta 
##                                                                                                           16105 
##                                          12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_9.fasta 
##                                                                                                           16282 
##                                           12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_1.fasta 
##                                                                                                           15716 
##                                          12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_10.fasta 
##                                                                                                           15455 
##                                           12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_2.fasta 
##                                                                                                           15781 
##                                           12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_3.fasta 
##                                                                                                           15799 
##                                           12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_4.fasta 
##                                                                                                           15581 
##                                           12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_5.fasta 
##                                                                                                           15704 
##                                           12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_6.fasta 
##                                                                                                           15553 
##                                           12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_7.fasta 
##                                                                                                           15589 
##                                           12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_8.fasta 
##                                                                                                           15583 
##                                           12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_9.fasta 
##                                                                                                           15626 
##                                                                        12s_v010_finalforTaxonomy.filtered.fasta 
##                                                                                                             240 
##   12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_10forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_1forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_2forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_3forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_4forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_5forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_6forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_7forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_8forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_9forTaxonomy.filtered.fasta 
##                                                                                                             240 
## 12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_10forTaxonomy.filtered.fasta 
##                                                                                                             240 
##  12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_1forTaxonomy.filtered.fasta 
##                                                                                                             240 
##  12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_2forTaxonomy.filtered.fasta 
##                                                                                                             240 
##  12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_3forTaxonomy.filtered.fasta 
##                                                                                                             240 
##  12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_4forTaxonomy.filtered.fasta 
##                                                                                                             240 
##  12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_5forTaxonomy.filtered.fasta 
##                                                                                                             240 
##  12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_6forTaxonomy.filtered.fasta 
##                                                                                                             240 
##  12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_7forTaxonomy.filtered.fasta 
##                                                                                                             240 
##  12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_8forTaxonomy.filtered.fasta 
##                                                                                                             240 
##  12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_9forTaxonomy.filtered.fasta 
##                                                                                                             240 
##  12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_10forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_1forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_2forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_3forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_4forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_5forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_6forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_7forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_8forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   12s_v010_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_9forTaxonomy.filtered.fasta 
##                                                                                                             240 
##                                                                                            12S_v10_HmmCut.fasta 
##                                                                                                            8714 
##                                                                                             16S_v04_final.fasta 
##                                                                                                           17674 
##                                             16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_1.fasta 
##                                                                                                           16374 
##                                            16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_10.fasta 
##                                                                                                           16360 
##                                             16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_2.fasta 
##                                                                                                           16735 
##                                             16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_3.fasta 
##                                                                                                           16567 
##                                             16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_4.fasta 
##                                                                                                           16776 
##                                             16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_5.fasta 
##                                                                                                           16689 
##                                             16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_6.fasta 
##                                                                                                           16330 
##                                             16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_7.fasta 
##                                                                                                           16841 
##                                             16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_8.fasta 
##                                                                                                           16165 
##                                             16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_9.fasta 
##                                                                                                           16285 
##                                           16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_1.fasta 
##                                                                                                           17050 
##                                          16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_10.fasta 
##                                                                                                           16867 
##                                           16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_2.fasta 
##                                                                                                           17165 
##                                           16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_3.fasta 
##                                                                                                           16830 
##                                           16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_4.fasta 
##                                                                                                           17045 
##                                           16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_5.fasta 
##                                                                                                           17369 
##                                           16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_6.fasta 
##                                                                                                           16953 
##                                           16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_7.fasta 
##                                                                                                           17104 
##                                           16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_8.fasta 
##                                                                                                           17262 
##                                           16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_9.fasta 
##                                                                                                           17159 
##                                            16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_1.fasta 
##                                                                                                           16172 
##                                           16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_10.fasta 
##                                                                                                           16327 
##                                            16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_2.fasta 
##                                                                                                           16294 
##                                            16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_3.fasta 
##                                                                                                           16191 
##                                            16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_4.fasta 
##                                                                                                           16435 
##                                            16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_5.fasta 
##                                                                                                           16174 
##                                            16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_6.fasta 
##                                                                                                           15983 
##                                            16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_7.fasta 
##                                                                                                           16145 
##                                            16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_8.fasta 
##                                                                                                           16010 
##                                            16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_9.fasta 
##                                                                                                           16118 
##                                                                         16S_v04_finalforTaxonomy.filtered.fasta 
##                                                                                                             240 
##    16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_10forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_1forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_2forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_3forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_4forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_5forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_6forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_7forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_8forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_9forTaxonomy.filtered.fasta 
##                                                                                                             240 
##  16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_10forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_1forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_2forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_3forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_4forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_5forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_6forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_7forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_8forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_9forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_10forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_1forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_2forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_3forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_4forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_5forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_6forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_7forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_8forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    16S_v04_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_9forTaxonomy.filtered.fasta 
##                                                                                                             240 
##                                                                                            16S_v04_HmmCut.fasta 
##                                                                                                            9590 
##                                                                                             c01_v03_final.fasta 
##                                                                                                           12609 
##                                             c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_1.fasta 
##                                                                                                           12286 
##                                            c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_10.fasta 
##                                                                                                           12907 
##                                             c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_2.fasta 
##                                                                                                           12333 
##                                             c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_3.fasta 
##                                                                                                           12919 
##                                             c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_4.fasta 
##                                                                                                           12107 
##                                             c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_5.fasta 
##                                                                                                           12272 
##                                             c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_6.fasta 
##                                                                                                           12223 
##                                             c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_7.fasta 
##                                                                                                           12673 
##                                             c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_8.fasta 
##                                                                                                           12229 
##                                             c01_v03_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_9.fasta 
##                                                                                                           12925 
##                                           c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_1.fasta 
##                                                                                                           12455 
##                                          c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_10.fasta 
##                                                                                                           12383 
##                                           c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_2.fasta 
##                                                                                                           12356 
##                                           c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_3.fasta 
##                                                                                                           12334 
##                                           c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_4.fasta 
##                                                                                                           12420 
##                                           c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_5.fasta 
##                                                                                                           12446 
##                                           c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_6.fasta 
##                                                                                                           12329 
##                                           c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_7.fasta 
##                                                                                                           12419 
##                                           c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_8.fasta 
##                                                                                                           12572 
##                                           c01_v03_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_9.fasta 
##                                                                                                           12499 
##                                            c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_1.fasta 
##                                                                                                           12989 
##                                           c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_10.fasta 
##                                                                                                           12948 
##                                            c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_2.fasta 
##                                                                                                           12809 
##                                            c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_3.fasta 
##                                                                                                           12053 
##                                            c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_4.fasta 
##                                                                                                           12010 
##                                            c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_5.fasta 
##                                                                                                           12513 
##                                            c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_6.fasta 
##                                                                                                           12803 
##                                            c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_7.fasta 
##                                                                                                           12641 
##                                            c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_8.fasta 
##                                                                                                           11991 
##                                            c01_v03_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_9.fasta 
##                                                                                                           12002 
##                                                                         c01_v03_finalforTaxonomy.filtered.fasta 
##                                                                                                             240 
##    c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_10forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_1forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_2forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_3forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_4forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_5forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_6forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_7forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_8forTaxonomy.filtered.fasta 
##                                                                                                             240 
##     c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_fifty_subset_9forTaxonomy.filtered.fasta 
##                                                                                                             240 
##  c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_10forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_1forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_2forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_3forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_4forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_5forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_6forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_7forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_8forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_seventy_subset_9forTaxonomy.filtered.fasta 
##                                                                                                             240 
##   c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_10forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_1forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_2forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_3forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_4forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_5forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_6forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_7forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_8forTaxonomy.filtered.fasta 
##                                                                                                             240 
##    c01_v03_finalforTaxonomy.filtered.fasta_Taxonomies.CountedFams.txt_thirty_subset_9forTaxonomy.filtered.fasta 
##                                                                                                             240 
##                                                                                            c01_v03_HmmCut.fasta 
##                                                                                                            6792
twelveS_data <- data |> filter(Subject == '12s_v010_final.fasta')
sixteenS_data <- data |> filter(Subject == '16S_v04_final.fasta')
table(twelveS_data$Query)
## 
##                                                                                KWest_16S_PooledTrue.fa 
##                                                                                                   4263 
##   make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_12S_Lulu_RESULTS_dada2_asv.fa 
##                                                                                                   1321 
## make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_12S_noLulu_RESULTS_dada2_asv.fa 
##                                                                                                   1321 
##   make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_16S_Lulu_RESULTS_dada2_asv.fa 
##                                                                                                   1095 
## make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_16S_noLulu_RESULTS_dada2_asv.fa 
##                                                                                                   1095 
##        make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_CO1_RESULTS_dada2_asv.fa 
##                                                                                                    968 
##                               make_12s_16s_simulated_reads_6-fakeGenes_GreenGenes_RESULTS_dada2_asv.fa 
##                                                                                                    990 
##                                   make_12s_16s_simulated_reads_6-fakeGenes_Random_RESULTS_dada2_asv.fa 
##                                                                                                   1000 
##                     make_12s_16s_simulated_reads_7-Lutjanids_Mock_runEDNAFlow_12S_RESULTS_dada2_asv.fa 
##                                                                                                    316 
##                     make_12s_16s_simulated_reads_7-Lutjanids_Mock_runEDNAFlow_16S_RESULTS_dada2_asv.fa 
##                                                                                                    299 
##                     make_12s_16s_simulated_reads_7-Lutjanids_Mock_runEDNAFLOW_CO1_RESULTS_dada2_asv.fa 
##                                                                                                    255 
##                           make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_12S_RESULTS_dada2_asv.fa 
##                                                                                                   1343 
##                           make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_16S_RESULTS_dada2_asv.fa 
##                                                                                                   1226 
##                           make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_CO1_RESULTS_dada2_asv.fa 
##                                                                                                   1098
table(sixteenS_data$Query)
## 
##                                                                                KWest_16S_PooledTrue.fa 
##                                                                                                   4928 
##   make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_12S_Lulu_RESULTS_dada2_asv.fa 
##                                                                                                   1230 
## make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_12S_noLulu_RESULTS_dada2_asv.fa 
##                                                                                                   1194 
##   make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_16S_Lulu_RESULTS_dada2_asv.fa 
##                                                                                                   1306 
## make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_16S_noLulu_RESULTS_dada2_asv.fa 
##                                                                                                   1306 
##        make_12s_16s_simulated_reads_5-BetterDatabaseARTSimulation_runEDNAFLOW_CO1_RESULTS_dada2_asv.fa 
##                                                                                                    968 
##                               make_12s_16s_simulated_reads_6-fakeGenes_GreenGenes_RESULTS_dada2_asv.fa 
##                                                                                                    990 
##                                   make_12s_16s_simulated_reads_6-fakeGenes_Random_RESULTS_dada2_asv.fa 
##                                                                                                   1000 
##                     make_12s_16s_simulated_reads_7-Lutjanids_Mock_runEDNAFlow_12S_RESULTS_dada2_asv.fa 
##                                                                                                    307 
##                     make_12s_16s_simulated_reads_7-Lutjanids_Mock_runEDNAFlow_16S_RESULTS_dada2_asv.fa 
##                                                                                                    341 
##                     make_12s_16s_simulated_reads_7-Lutjanids_Mock_runEDNAFLOW_CO1_RESULTS_dada2_asv.fa 
##                                                                                                    255 
##                           make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_12S_RESULTS_dada2_asv.fa 
##                                                                                                   1274 
##                           make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_16S_RESULTS_dada2_asv.fa 
##                                                                                                   1477 
##                           make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_CO1_RESULTS_dada2_asv.fa 
##                                                                                                   1098
table(sixteenS_data$Subject)
## 
## 16S_v04_final.fasta 
##               17674
twelveS_data_vs_12S_100 <- twelveS_data |> 
  filter(Query == 'make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_12S_RESULTS_dada2_asv.fa')
sixteenS_data_vs_16S_100 <- sixteenS_data |> 
  filter(Query == 'make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_16S_RESULTS_dada2_asv.fa' )
twelveS_data_vs_12S_100 |> select(Type, species) |> filter(species != 'dropped' &
                                                             !is.na(species)) |>
  group_by(Type) |> count(species) |> summarise(n = n()) |>
  ggplot(aes(x = Type, y = n, fill = n)) + geom_col() + coord_flip() + 
  theme_minimal() + 
  ylab('Count') + 
  ggtitle('12S: Species-level hits per classifier')

twelveS_data_vs_12S_100 |> select(Type, genus) |> filter(genus != 'dropped' &
                                                             !is.na(genus)) |>
  group_by(Type) |> count(genus) |> summarise(n = n()) |>
  ggplot(aes(x = Type, y = n, fill = n)) + geom_col() + coord_flip() + 
  theme_minimal() + 
  ylab('Count') + 
  ggtitle('12S: Genus-level hits per classifier')

twelveS_truth <- truth |> filter(Query == 'make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_12S_RESULTS_dada2_asv.fa') |> select(OTU, family, species) |> rename(True_OTU = OTU, True_family = family, True_species = species)
head(twelveS_truth)
## # A tibble: 6 × 3
##   True_OTU True_family  True_species            
##   <chr>    <chr>        <chr>                   
## 1 ASV_1    Syngnathidae Phyllopteryx taeniolatus
## 2 ASV_2    Syngnathidae Phycodurus eques        
## 3 ASV_3    Syngnathidae Phyllopteryx taeniolatus
## 4 ASV_4    Labridae     dropped                 
## 5 ASV_5    Alopiidae    Isurus oxyrinchus       
## 6 ASV_6    Carangidae   Seriola lalandi
twelveS_data_vs_12S_100 |> left_join(twelveS_truth, by = c('OTU' = 'True_OTU')) |>
  mutate(Correct = True_species == species) |> 
  filter(species != 'dropped' & !is.na(species)) |> 
  group_by(Type) |> count(Correct) |> 
  ggplot(aes(x = fct_rev(fct_reorder2(Type, Correct, n)), fill = Correct, y = n))+ geom_col() +
  coord_flip() + theme_minimal() + xlab('Type') + 
  ggtitle('12S: Correct and incorrect species-level classifications (absolute)') +
  scale_fill_manual(values = c("#E69F00", "#56B4E9", "#009E73",
          "#F0E442", "#0072B2", "#D55E00", "#CC79A7"))

cols <- c('Correct species' = "#009E73", 'Correct genus'="#56B4E9", 'Correct family' = "#0072B2", 'Incorrect family' = "#E69F00", 'Incorrect genus'="#F0E442", 'Incorrect species'="#D55E00", 'No hit'= "#CC79A7")
twelve_s_relative_plot <- twelveS_data_vs_12S_100 |> left_join(twelveS_truth, by = c('OTU' = 'True_OTU')) |>
  separate(True_species, into = c('True_Genus', 'True_Epiteth'), remove = FALSE)|> 
  mutate(species = na_if(species, 'dropped')) |> 
  mutate(genus = na_if(genus, 'dropped')) |> 

  mutate(CorrectSpecies = case_when(!is.na(species) & True_species == species ~ 'Correct species',
                              !is.na(species) & True_species != species ~ 'Incorrect species',
                              !is.na(genus) & !is.na(True_Genus) & True_Genus == genus ~ 'Correct genus',
                              !is.na(genus) & !is.na(True_Genus) & True_Genus != genus ~ 'Incorrect genus',
                              !is.na(family) & True_family == family ~ 'Correct family',
                              !is.na(family) & True_family != family ~ 'Incorrect family',
                              TRUE ~ NA)) |> 
  group_by(Type) |>
  count(CorrectSpecies) |> 
  mutate(total = sum(n, na.rm=TRUE)) |> 
  mutate(missing = 102 - total) |> 
  group_modify(~ add_row(.x)) |> 
  group_modify(~ {
    .x |> mutate(new_col= max(missing, na.rm=TRUE)) |> 
      mutate(n = case_when(is.na(CorrectSpecies) & is.na(missing) ~ new_col,
                                  TRUE ~ n)) |> 
      select(-new_col)
  } ) |> 
  mutate(total = 102) |> 
  mutate(perc = n / total * 100) |> 
  mutate(CorrectSpecies = replace_na(CorrectSpecies, 'No hit')) |> 
  mutate(CorrectSpecies = factor(CorrectSpecies, rev(c('Correct species', 'Correct genus', 'Correct family', 'Incorrect family', 'Incorrect genus', 'Incorrect species', 'No hit')))) |> 
  ggplot(aes(x = fct_rev(fct_reorder2(Type, CorrectSpecies, n)), fill = CorrectSpecies, y = perc))+ 
  geom_col() +
  coord_flip() + 
  theme_minimal() + 
  ylab('Percentage') + xlab('Type') +
  ggtitle('12S: Correct and incorrect species-level classifications (relative)') +
  scale_fill_manual(name = 'Outcome', values = cols, breaks=names(cols))
twelve_s_relative_plot

### Calculate Upset-based species sightings

type_list <- twelveS_data_vs_12S_100 |> select(Type, species) |> unique() |> filter(!is.na(species) & species != 'dropped') |> 
  group_by(species) |> 
  summarize('Type' = list(Type))

a <- type_list |>
  ggplot(aes(x = Type)) + 
  geom_bar() +
  scale_x_upset(n_intersections = 12) +
  ggtitle('12S: Shared species') +
  ylab('Species')
a

type_list <- twelveS_data_vs_12S_100 |> left_join(twelveS_truth, by = c('OTU' = 'True_OTU')) |>
  filter(species == True_species) |> 
  filter(species != 'dropped' & !is.na(species)) |> 
  select(Type, species) |> unique() |> 
  group_by(species) |> 
  summarize('Type' = list(Type))

b <- type_list |>
  ggplot(aes(x = Type)) + 
  geom_bar() +
  scale_x_upset(n_intersections = 12) +
  ggtitle('12S: Shared correct species') +
  ylab('Species')
b

type_list <- twelveS_data_vs_12S_100 |> left_join(twelveS_truth, by = c('OTU' = 'True_OTU')) |>
  filter(species != True_species) |> 
  filter(species != 'dropped' & !is.na(species)) |> 
  select(Type, species) |> unique() |> 
  group_by(species) |> 
  summarize('Type' = list(Type))

c <- type_list |>
  ggplot(aes(x = Type)) + 
  geom_bar() +
  scale_x_upset(n_intersections = 12) +
  ggtitle('12S: Shared incorrect species') +
  ylab('Species')
c

a + b + c & ylim(c(0, 30)) & 
  theme(
  # Hide panel borders and remove grid lines
  panel.border = element_blank(),
  panel.grid.major = element_blank(),
  panel.background = element_blank(),
  panel.grid.minor = element_blank(),
  #panel.grid.major.y = element_line(),
  # Change axis line
  axis.line = element_line(colour = "black")
  )

Calculate FP/TP/TN/FN

add_scores <- function(twelveS_data_vs_12S_100_with_MaxTruth, twelveS_truth ) {
  twelveS_data_vs_12S_100_with_MaxTruth|> left_join(twelveS_truth, by = c('OTU' = 'True_OTU')) |>
  separate(True_species, into = c('True_Genus', 'True_Epiteth'), remove = FALSE)|> 
  mutate(species = na_if(species, 'dropped')) |> 
  mutate(genus = na_if(genus, 'dropped')) |> 

  mutate(CorrectSpecies = case_when(!is.na(species) & True_species == species ~ 'Correct species',
                              !is.na(species) & True_species != species ~ 'Incorrect species',
                              !is.na(genus) & !is.na(True_Genus) & True_Genus == genus ~ 'Correct genus',
                              !is.na(genus) & !is.na(True_Genus) & True_Genus != genus ~ 'Incorrect genus',
                              !is.na(family) & True_family == family ~ 'Correct family',
                              !is.na(family) & True_family != family ~ 'Incorrect family',
                              TRUE ~ NA)) |> 
  group_by(Type) |>
  summarise(TP = sum(str_detect(CorrectSpecies, pattern='Correct species'), na.rm=TRUE),
            FP = sum(str_detect(CorrectSpecies, pattern = 'Incorrect species'), na.rm=TRUE),
            TN = sum(str_detect(replace_na(CorrectSpecies,'NA'), pattern='NA') & is.na(True_species), na.rm=TRUE),
            FN = sum(is.na(species) & !is.na(True_species))) |> 
  mutate(sums = TP + FP + TN + FN) |> 
  mutate(missing = 102 - sums) |> 
  mutate(FN = FN + missing) |> 
  mutate(sums = TP + FP + TN + FN) |> 
  select(-c(missing, sums))
}
scores <- add_scores(twelveS_data_vs_12S_100, twelveS_truth)
scores <- scores |> rowwise() |> mutate(recall = recall(TP, FN), precision = precision(TP, FP),
                              f1 = f1(precision, recall), f0.5 = f0.5(precision, recall), accuracy = accuracy(TP, FP, FN, TN))
scores
## # A tibble: 14 × 10
## # Rowwise: 
##    Type           TP    FP    TN    FN recall precision    f1  f0.5 accuracy
##    <chr>       <int> <int> <int> <dbl>  <dbl>     <dbl> <dbl> <dbl>    <dbl>
##  1 BLAST100       57     2     0    43 0.57       0.966 0.717 0.848   0.559 
##  2 BLAST97        59     8     0    35 0.628      0.881 0.733 0.815   0.578 
##  3 CustomNBC      53    18     0    31 0.631      0.746 0.684 0.720   0.520 
##  4 DADA2Spec      57     3     0    42 0.576      0.95  0.717 0.841   0.559 
##  5 DADA2Tax        7    26     0    69 0.0921     0.212 0.128 0.168   0.0686
##  6 Kraken_0.0     60    12     0    30 0.667      0.833 0.741 0.794   0.588 
##  7 Kraken_0.05    57     9     0    36 0.613      0.864 0.717 0.798   0.559 
##  8 Kraken_0.1     53     5     0    44 0.546      0.914 0.684 0.805   0.520 
##  9 MMSeqs2_100    58     2     0    42 0.58       0.967 0.725 0.853   0.569 
## 10 MMSeqs2_97     64     7     0    31 0.674      0.901 0.771 0.844   0.627 
## 11 Metabuli       65    11     0    26 0.714      0.855 0.778 0.823   0.637 
## 12 Mothur         42    22     0    38 0.525      0.656 0.583 0.625   0.412 
## 13 Qiime2         31    18     0    53 0.369      0.633 0.466 0.554   0.304 
## 14 VSEARCH        52    17     0    33 0.612      0.754 0.675 0.720   0.510
twelveS_scoreS_plot <- scores |> select(-c(TP, FP, FN, TN)) |> pivot_longer(-Type, names_to='Score') |>  ggplot(aes(x = fct_rev(fct_reorder(Type, value)), y = value, group=Score, color = Score, fill =Score)) + geom_line() + ylim(c(0, 1))  + theme_minimal_hgrid()+ theme(axis.text.x = element_text( angle = 45, hjust = 1)) + ylab('Score') + xlab('Tool') + ggtitle('12S scores')
twelveS_scoreS_plot

Scores heatmap

Let’s also make a heatmap from that

b <- scores$Type
m <- scores |> select(-Type) |> select(accuracy, recall, precision, f1, f0.5) |> as.matrix()
rownames(m) <- b
pheatmap(m, cluster_cols=FALSE, display_numbers = TRUE,
         color = colorRampPalette(brewer.pal(n = 7, name =
  "RdYlGn"))(100))

b <- scores$Type
m <- scores |> select(-Type) |> select(TP, FP, FN) |> as.matrix()
rownames(m) <- b
pheatmap(m, cluster_cols=FALSE, display_numbers = TRUE,
         color = colorRampPalette(brewer.pal(n = 7, name =
  "RdYlGn"))(100))

Metaclassifier

table(twelveS_data_vs_12S_100$Type)
## 
##    BLAST100     BLAST97   CustomNBC   DADA2Spec    DADA2Tax  Kraken_0.0 
##          79          93         102         102         102         102 
## Kraken_0.05  Kraken_0.1    Metabuli MMSeqs2_100  MMSeqs2_97      Mothur 
##         102         102         102         102         102         102 
##      Qiime2     VSEARCH 
##          49         102

First, we count the per-OTU species hits

twelveS_data_vs_12S_100_maxCount <- twelveS_data_vs_12S_100 |>  
  mutate(species = na_if(species, 'dropped')) |> 
  filter(!is.na(species)) |> 
  #filter(! Type %in% c('Mothur', 'VSEARCH', 'Kraken_0.2', 'Qiime2', 'Metabuli', 'NBC', 'BLAST97', 'Kraken_0.0', 'Kraken_0.1')) |> 
  group_by(Query, Subject, OTU) |> 
  count(species) |> 
  # double check the truth
  #left_join(twelveS_truth, by = c('OTU' = 'True_OTU')) |> 
  #mutate(Truth = True_species == species) |> 
  # pull out the per-group highest n
  filter( n > 4) |> 
  slice_max(n, n=1, with_ties = FALSE) |> 
  mutate(Type = 'MaxCount', .before = 'Query') |> 
  select(-n)
twelveS_data_vs_12S_100_maxCount
## # A tibble: 75 × 5
## # Groups:   Query, Subject, OTU [75]
##    Type     Query                                          Subject OTU   species
##    <chr>    <chr>                                          <chr>   <chr> <chr>  
##  1 MaxCount make_12s_16s_simulated_reads_8-Rottnest_runED… 12s_v0… ASV_1 Phyllo…
##  2 MaxCount make_12s_16s_simulated_reads_8-Rottnest_runED… 12s_v0… ASV_… Carcha…
##  3 MaxCount make_12s_16s_simulated_reads_8-Rottnest_runED… 12s_v0… ASV_… Isurus…
##  4 MaxCount make_12s_16s_simulated_reads_8-Rottnest_runED… 12s_v0… ASV_… Acanth…
##  5 MaxCount make_12s_16s_simulated_reads_8-Rottnest_runED… 12s_v0… ASV_… Pseudo…
##  6 MaxCount make_12s_16s_simulated_reads_8-Rottnest_runED… 12s_v0… ASV_… Carcha…
##  7 MaxCount make_12s_16s_simulated_reads_8-Rottnest_runED… 12s_v0… ASV_… Parupe…
##  8 MaxCount make_12s_16s_simulated_reads_8-Rottnest_runED… 12s_v0… ASV_… Sphyrn…
##  9 MaxCount make_12s_16s_simulated_reads_8-Rottnest_runED… 12s_v0… ASV_… Parupe…
## 10 MaxCount make_12s_16s_simulated_reads_8-Rottnest_runED… 12s_v0… ASV_… Chaeto…
## # ℹ 65 more rows
twelveS_data_vs_12S_100_with_MaxTruth <- twelveS_data_vs_12S_100 |> 
  bind_rows(twelveS_data_vs_12S_100_maxCount) #|>  
  #filter(! Type %in% c('Mothur', 'VSEARCH', 'Kraken_0.2', 'Qiime2', 'Metabuli', 'NBC', 'BLAST97', 'Kraken_0.0', 'Kraken_0.1')) 
maxTruth_scores <-  add_scores(twelveS_data_vs_12S_100_with_MaxTruth, twelveS_truth )
maxTruth_scores <- maxTruth_scores |> rowwise() |> mutate(recall = recall(TP, FN), precision = precision(TP, FP),
                              f1 = f1(precision, recall), f0.5 = f0.5(precision, recall), accuracy = accuracy(TP, FP, FN, TN))
maxTruth_scoreS_plot <- maxTruth_scores |> select(-c(TP, FP, FN, TN)) |> pivot_longer(-Type, names_to='Score') |>  ggplot(aes(x = fct_rev(fct_reorder(Type, value)), y = value, group=Score, color = Score, fill =Score)) + geom_line() + ylim(c(0, 1))  + theme_minimal_hgrid()+ theme(axis.text.x = element_text( angle = 45, hjust = 1)) + ylab('Score') + xlab('Tool') + geom_point() + ggtitle('12S scores')
maxTruth_scoreS_plot

With the Wadjemup data MaxCount performs slightly better! But still not as good as MMSeqs2….

16S

sixteenS_truth <- truth |> filter(Query == 'make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_16S_RESULTS_dada2_asv.fa') |> select(OTU, family, species) |> rename(True_OTU = OTU, True_family = family, True_species = species)
tail(sixteenS_truth)
## # A tibble: 6 × 3
##   True_OTU True_family    True_species            
##   <chr>    <chr>          <chr>                   
## 1 ASV_106  Alopiidae      Isurus oxyrinchus       
## 2 ASV_107  Carangidae     Seriola dumerili        
## 3 ASV_108  Carcharhinidae Sphyrna lewini          
## 4 ASV_109  Scombridae     Auxis thazard           
## 5 ASV_110  Syngnathidae   Phycodurus eques        
## 6 ASV_111  Syngnathidae   Phyllopteryx taeniolatus
sixteenS_relative_plot <- sixteenS_data_vs_16S_100 |> left_join(sixteenS_truth, by = c('OTU' = 'True_OTU')) |>
 separate(True_species, into = c('True_Genus', 'True_Epiteth'), remove = FALSE)|> 
  mutate(species = na_if(species, 'dropped')) |> 
  mutate(genus = na_if(genus, 'dropped')) |> 

  mutate(CorrectSpecies = case_when(!is.na(species) & True_species == species ~ 'Correct species',
                              !is.na(species) & True_species != species ~ 'Incorrect species',
                              !is.na(genus) & !is.na(True_Genus) & True_Genus == genus ~ 'Correct genus',
                              !is.na(genus) & !is.na(True_Genus) & True_Genus != genus ~ 'Incorrect genus',
                              !is.na(family) & True_family == family ~ 'Correct family',
                              !is.na(family) & True_family != family ~ 'Incorrect family',
                              TRUE ~ NA)) |> 
  group_by(Type) |> 
  count(CorrectSpecies) |> 
  mutate(total = sum(n)) |> 
  mutate(missing = 112 - total) |> 
  group_modify(~ add_row(.x)) |> 
  group_modify(~ {
    .x |> mutate(new_col= max(missing, na.rm=TRUE)) |> 
      mutate(n = case_when(is.na(CorrectSpecies) & is.na(missing) ~ new_col,
                                  TRUE ~ n)) |> 
      select(-new_col)
  } ) |> 
  mutate(total = 112) |> 
  mutate(perc = n / total * 100) |> 
  mutate(CorrectSpecies = replace_na(CorrectSpecies, 'No hit')) |> 
  mutate(CorrectSpecies = factor(CorrectSpecies, rev(c('Correct species', 'Correct genus', 'Correct family', 'Incorrect family', 'Incorrect genus', 'Incorrect species', 'No hit')))) |> 
  tidyr::complete(CorrectSpecies, fill = list(n=0, total = 112, missing = NA, perc = 0)) |>  
  ggplot(aes(x = fct_rev(fct_reorder2(Type, CorrectSpecies, n)), fill = CorrectSpecies, y = perc))+ 
  geom_col() +
  coord_flip() + 
  theme_minimal() + 
  ylab('Percentage') + xlab('Type') +
  ggtitle('16S: Correct and incorrect species-level classifications (relative)') +
  scale_fill_manual(name = 'Outcome', values = cols, breaks=names(cols))
sixteenS_relative_plot 

Calculate scores

scores <- add_scores(sixteenS_data_vs_16S_100, sixteenS_truth)
scores
## # A tibble: 14 × 5
##    Type           TP    FP    TN    FN
##    <chr>       <int> <int> <int> <dbl>
##  1 BLAST100       53     3     0    46
##  2 BLAST97        62     6     2    32
##  3 CustomNBC      56    18     2    26
##  4 DADA2Spec      52     4     2    44
##  5 DADA2Tax        8    40     2    52
##  6 Kraken_0.0     70    16     2    14
##  7 Kraken_0.05    69    11     2    20
##  8 Kraken_0.1     66     8     2    26
##  9 MMSeqs2_100    54     3     2    43
## 10 MMSeqs2_97     69     8     2    23
## 11 Metabuli       69    18     2    13
## 12 Mothur         61    27     2    12
## 13 Qiime2         56    20     1    25
## 14 VSEARCH        68    11     2    21
scores <- scores |> rowwise() |> mutate(recall = recall(TP, FN), precision = precision(TP, FP),
                              f1 = f1(precision, recall), f0.5 = f0.5(precision, recall), accuracy = accuracy(TP, FP, FN, TN))
scores
## # A tibble: 14 × 10
## # Rowwise: 
##    Type           TP    FP    TN    FN recall precision    f1  f0.5 accuracy
##    <chr>       <int> <int> <int> <dbl>  <dbl>     <dbl> <dbl> <dbl>    <dbl>
##  1 BLAST100       53     3     0    46  0.535     0.946 0.684 0.820   0.520 
##  2 BLAST97        62     6     2    32  0.660     0.912 0.765 0.847   0.627 
##  3 CustomNBC      56    18     2    26  0.683     0.757 0.718 0.741   0.569 
##  4 DADA2Spec      52     4     2    44  0.542     0.929 0.684 0.813   0.529 
##  5 DADA2Tax        8    40     2    52  0.133     0.167 0.148 0.159   0.0980
##  6 Kraken_0.0     70    16     2    14  0.833     0.814 0.824 0.818   0.706 
##  7 Kraken_0.05    69    11     2    20  0.775     0.862 0.817 0.844   0.696 
##  8 Kraken_0.1     66     8     2    26  0.717     0.892 0.795 0.851   0.667 
##  9 MMSeqs2_100    54     3     2    43  0.557     0.947 0.701 0.831   0.549 
## 10 MMSeqs2_97     69     8     2    23  0.75      0.896 0.817 0.862   0.696 
## 11 Metabuli       69    18     2    13  0.841     0.793 0.817 0.802   0.696 
## 12 Mothur         61    27     2    12  0.836     0.693 0.758 0.718   0.618 
## 13 Qiime2         56    20     1    25  0.691     0.737 0.713 0.727   0.559 
## 14 VSEARCH        68    11     2    21  0.764     0.861 0.810 0.840   0.686
sixteenS_score_plot <- scores |> select(-c(TP, FP, FN, TN)) |> pivot_longer(-Type, names_to='Score') |>  ggplot(aes(x = fct_rev(fct_reorder(Type, value)), y = value, group=Score, color = Score, fill =Score)) + geom_line() + ylim(c(0, 1))  + theme_minimal_hgrid() + theme(axis.text.x = element_text( angle = 45, hjust = 1)) + ylab('Score') + xlab('Tool') +  ggtitle('16S scores')
sixteenS_score_plot 

b <- scores$Type
m <- scores |> select(-Type) |> select(accuracy, recall, precision, f1, f0.5) |> as.matrix()
rownames(m) <- b
pheatmap(m, cluster_cols=FALSE, display_numbers = TRUE,
         color = colorRampPalette(brewer.pal(n = 7, name =
  "RdYlGn"))(100))

b <- scores$Type
m <- scores |> select(-Type) |> select(TP, FP, FN) |> as.matrix()
rownames(m) <- b
pheatmap(m, cluster_cols=FALSE, display_numbers = TRUE,
         color = colorRampPalette(brewer.pal(n = 7, name =
  "RdYlGn"))(100))

### Calculate Upset-based species sightings

type_list <- sixteenS_data_vs_16S_100  |> select(Type, species) |> unique() |> filter(!is.na(species) & species != 'dropped') |> 
  group_by(species) |> 
  summarize('Type' = list(Type))

a <- type_list |>
  ggplot(aes(x = Type)) + 
  geom_bar() +
  scale_x_upset(n_intersections = 12) +
  ggtitle('16S: Shared species') +
  ylab('Species')
a

type_list <- sixteenS_data_vs_16S_100 |> left_join(sixteenS_truth,  by = c('OTU' = 'True_OTU')) |>
  filter(species == True_species) |> 
  filter(species != 'dropped' & !is.na(species)) |> 
  select(Type, species) |> unique() |> 
  group_by(species) |> 
  summarize('Type' = list(Type))

b <- type_list |>
  ggplot(aes(x = Type)) + 
  geom_bar() +
  scale_x_upset(n_intersections = 12) +
  ggtitle('16S: Shared correct species') +
  ylab('Species')
b

type_list <- sixteenS_data_vs_16S_100 |> left_join(sixteenS_truth, by = c('OTU' = 'True_OTU')) |>
  filter(species != True_species) |> 
  filter(species != 'dropped' & !is.na(species)) |> 
  select(Type, species) |> unique() |> 
  group_by(species) |> 
  summarize('Type' = list(Type))

c <- type_list |>
  ggplot(aes(x = Type)) + 
  geom_bar() +
  scale_x_upset(n_intersections = 12) +
  ggtitle('16S: Shared incorrect species') +
  ylab('Species')
c

a + b + c & ylim(c(0, 20)) & 
  theme(
  # Hide panel borders and remove grid lines
  panel.border = element_blank(),
  panel.grid.major = element_blank(),
  panel.background = element_blank(),
  panel.grid.minor = element_blank(),
  #panel.grid.major.y = element_line(),
  # Change axis line
  axis.line = element_line(colour = "black")
  )

16S/12S relative plots

sixteenS_relative_plot / twelve_s_relative_plot

Let’s make without titles, but with a/b

(sixteenS_relative_plot + ggtitle('') + ylab(''))/ (twelve_s_relative_plot + ggtitle('')) + 
  plot_annotation(tag_levels = c('A','B')) +
  plot_layout(guides = 'collect')

(sixteenS_score_plot +geom_point() + theme(axis.title.x = element_blank()))/ (twelveS_scoreS_plot + geom_point())

12S exclusion databases

twelve_exclusions <- data |> filter(str_starts(Subject, '12s_v010_final.fasta_Taxonomies.CountedFams.txt_')) |> 
  filter(Query == 'make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_12S_RESULTS_dada2_asv.fa')
table(twelve_exclusions$Subject)
## 
##    12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_1.fasta 
##                                                                    1242 
##   12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_10.fasta 
##                                                                    1248 
##    12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_2.fasta 
##                                                                    1259 
##    12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_3.fasta 
##                                                                    1234 
##    12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_4.fasta 
##                                                                    1213 
##    12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_5.fasta 
##                                                                    1244 
##    12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_6.fasta 
##                                                                    1232 
##    12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_7.fasta 
##                                                                    1230 
##    12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_8.fasta 
##                                                                    1267 
##    12s_v010_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_9.fasta 
##                                                                    1246 
##  12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_1.fasta 
##                                                                    1274 
## 12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_10.fasta 
##                                                                    1265 
##  12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_2.fasta 
##                                                                    1196 
##  12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_3.fasta 
##                                                                    1213 
##  12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_4.fasta 
##                                                                    1276 
##  12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_5.fasta 
##                                                                    1296 
##  12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_6.fasta 
##                                                                    1250 
##  12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_7.fasta 
##                                                                    1299 
##  12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_8.fasta 
##                                                                    1257 
##  12s_v010_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_9.fasta 
##                                                                    1266 
##   12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_1.fasta 
##                                                                    1190 
##  12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_10.fasta 
##                                                                    1179 
##   12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_2.fasta 
##                                                                    1170 
##   12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_3.fasta 
##                                                                    1166 
##   12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_4.fasta 
##                                                                    1146 
##   12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_5.fasta 
##                                                                    1193 
##   12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_6.fasta 
##                                                                    1159 
##   12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_7.fasta 
##                                                                    1157 
##   12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_8.fasta 
##                                                                    1183 
##   12s_v010_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_9.fasta 
##                                                                    1216
twelve_exclusions_split <- twelve_exclusions |> separate(Subject, into = c('before', 'hit'), sep='.txt_') |> 
  # get rid of leftover non-subsetted databases
  filter(!is.na(hit)) |> 
  separate(hit, into=c('Database', 'after'), sep='_subset')
twelve_exclusions_split_averaged <- twelve_exclusions_split |> left_join(twelveS_truth, by = c('OTU' = 'True_OTU')) |>
  separate(True_species, into = c('True_Genus', 'True_Epiteth'), remove = FALSE)|> 
  mutate(species = na_if(species, 'dropped')) |> 
  mutate(genus = na_if(genus, 'dropped')) |> 

  mutate(CorrectSpecies = case_when(!is.na(species) & True_species == species ~ 'Correct species',
                              !is.na(species) & True_species != species ~ 'Incorrect species',
                              !is.na(genus) & !is.na(True_Genus) & True_Genus == genus ~ 'Correct genus',
                              !is.na(genus) & !is.na(True_Genus) & True_Genus != genus ~ 'Incorrect genus',
                              !is.na(family) & True_family == family ~ 'Correct family',
                              !is.na(family) & True_family != family ~ 'Incorrect family',
                              TRUE ~ NA)) |> 
  group_by(Type, Database, after) |>
  summarise(TP = sum(str_detect(CorrectSpecies, pattern='Correct species'), na.rm=TRUE),
            FP = sum(str_detect(CorrectSpecies, pattern = 'Incorrect species'), na.rm=TRUE),
            TN = sum(str_detect(replace_na(CorrectSpecies,'NA'), pattern='NA') & is.na(True_species), na.rm=TRUE),
            FN = sum(is.na(species) & !is.na(True_species))) |> 
  mutate(sums = TP + FP + TN + FN) |> 
  mutate(missing = 102 - sums) |> 
  mutate(FN = FN + missing) |> 
  mutate(sums = TP + FP + TN + FN) |> 
  select(-c(missing, sums)) |> 
  group_by(Type, Database) |> 
  summarise(mean_TP = mean(TP),
            mean_FP = mean(FP),
            mean_TN = mean(TN),
            mean_FN = mean(FN)) |> 
  rowwise() |> 
  mutate(recall = recall(mean_TP, mean_FN), 
         precision = precision(mean_TP, mean_FP),
         f1 = f1(precision, recall),
         f0.5 = f0.5(precision, recall),
         accuracy = accuracy(mean_TP, mean_FP, mean_FN, mean_TN)) 
## `summarise()` has grouped output by 'Type', 'Database'. You can override using
## the `.groups` argument.
## `summarise()` has grouped output by 'Type'. You can override using the
## `.groups` argument.
twelve_exclusions_split_averaged <- twelve_exclusions_split_averaged |> 
  mutate(Database = case_when ( Database == 'fifty' ~ '50%',
                               Database == 'seventy' ~ '70%',
                               Database == 'thirty' ~ '30%',
                               TRUE ~ Database))
f1_pl <- twelve_exclusions_split_averaged |> 
  ggplot(aes(x = Type, y = f1, group = Database, color = Database, fill = Database)) + 
  geom_point() +
  geom_line() +
  theme_minimal()
f0.5_pl <- twelve_exclusions_split_averaged |> 
  ggplot(aes(x = Type, y = f0.5, group = Database, color = Database, fill = Database)) + 
  geom_point() +
  geom_line() +
  theme_minimal()
precision_pl <- twelve_exclusions_split_averaged |> 
  ggplot(aes(x = Type, y = precision, group = Database, color = Database, fill = Database)) + 
  geom_point() +
  geom_line() +
  theme_minimal()
recall_pl <- twelve_exclusions_split_averaged |> 
  ggplot(aes(x = Type, y = recall, group = Database, color = Database, fill = Database)) + 
  geom_point() +
  geom_line() +
  theme_minimal()
(f1_pl / f0.5_pl / precision_pl / recall_pl) +  plot_layout(guides = 'collect')

Lets zero in on the precision and make boxplots with jitter dots

un_summarised_twelve <- twelve_exclusions_split |> left_join(twelveS_truth, by = c('OTU' = 'True_OTU')) |>
  separate(True_species, into = c('True_Genus', 'True_Epiteth'), remove = FALSE)|> 
  mutate(species = na_if(species, 'dropped')) |> 
  mutate(genus = na_if(genus, 'dropped')) |> 

  mutate(CorrectSpecies = case_when(!is.na(species) & True_species == species ~ 'Correct species',
                              !is.na(species) & True_species != species ~ 'Incorrect species',
                              !is.na(genus) & !is.na(True_Genus) & True_Genus == genus ~ 'Correct genus',
                              !is.na(genus) & !is.na(True_Genus) & True_Genus != genus ~ 'Incorrect genus',
                              !is.na(family) & True_family == family ~ 'Correct family',
                              !is.na(family) & True_family != family ~ 'Incorrect family',
                              TRUE ~ NA)) |> 
  group_by(Type, Database, after) |>
  summarise(TP = sum(str_detect(CorrectSpecies, pattern='Correct species'), na.rm=TRUE),
            FP = sum(str_detect(CorrectSpecies, pattern = 'Incorrect species'), na.rm=TRUE),
            TN = sum(str_detect(replace_na(CorrectSpecies,'NA'), pattern='NA') & is.na(True_species), na.rm=TRUE),
            FN = sum(is.na(species) & !is.na(True_species))) |> 
  mutate(sums = TP + FP + TN + FN) |> 
  mutate(missing = 102 - sums) |> 
  mutate(FN = FN + missing) |> 
  mutate(sums = TP + FP + TN + FN) |> 
  select(-c(missing, sums)) |> 
  rowwise() |> 
  mutate(recall = recall(TP, FN), 
         precision = precision(TP, FP),
         f1 = f1(precision, recall),
         f0.5 = f0.5(precision, recall),
         accuracy = accuracy(TP, FP, FN, TN)) |> 
  mutate(Database = case_when ( Database == 'fifty' ~ '50%',
                               Database == 'seventy' ~ '70%',
                               Database == 'thirty' ~ '30%',
                               TRUE ~ Database))
## `summarise()` has grouped output by 'Type', 'Database'. You can override using
## the `.groups` argument.
un_summarised_twelve |> group_by(Type, Database) |> mutate(best = max(mean(precision))) |>
  ggplot(aes(x = fct_reorder(Type, best), group = interaction(Type, Database),  color=Database, y = precision)) +
  geom_boxplot(outlier.shape = NA) +
  coord_flip() +
  theme_minimal() +
  xlab('Type') +
  ylab('Precision') +
  geom_point(position = position_jitterdodge(), alpha=0.5)

un_summarised_twelve  |> group_by(Type, Database) |> mutate(best = max(mean(f0.5))) |>
  ggplot(aes(x = fct_reorder(Type, best), group = interaction(Type, Database), color=Database, y = f0.5)) +
  geom_boxplot(outlier.shape = NA) + 
  coord_flip() + 
  theme_minimal() + 
  xlab('Type') +
  ylab('f0.5') +
  ylim(c(0, 1)) +
  geom_point(position = position_jitterdodge(), alpha=0.5)

un_summarised_twelve  |> group_by(Type, Database) |> mutate(best = max(mean(recall))) |>
  ggplot(aes(x = fct_reorder(Type, best), group = interaction(Type, Database), color=Database, y = recall)) +
  geom_boxplot(outlier.shape = NA) + 
  coord_flip() + 
  theme_minimal() + 
  xlab('Type') +
  ylab('f0.5') +
  ylim(c(0, 1)) +
  geom_point(position = position_jitterdodge(), alpha=0.5)

un_summarised_twelve  |> 
  filter(Type %in% c('BLAST100', 'Kraken_0.0', 'Kraken_0.05', 'Qiime2', 'TNT')) |> 
  ggplot(aes(x=Database, y = precision, fill=Type)) + #fill=factor(Database, levels=c('30%','50%','70%')))) + 
  geom_boxplot() +
  labs(fill='Type') + 
  ylab('Precision') + 
  theme_minimal()

false_positives <- un_summarised_twelve  |> 
  filter(Type %in% c('BLAST100', 'Kraken_0.0', 'Kraken_0.05', 'Kraken_0.1', 'MMSeqs2', 'TNT')) |> 
  ggplot(aes(x=Database, y = FP/102*100, fill=Type)) + #fill=factor(Database, levels=c('30%','50%','70%')))) + 
  geom_boxplot(outlier.shape=NA) +
  labs(fill='Type') + 
  ylab('False positives (%)') + 
  geom_point(aes(color=Type), 
             position = position_jitterdodge(jitter.width = 0.2), 
             alpha=0.5,
             show.legend = FALSE)+
  theme_minimal()
false_positives

true_positives <- un_summarised_twelve  |> 
  filter(Type %in% c('BLAST100', 'Kraken_0.0', 'Kraken_0.05', 'Kraken_0.1', 'MMSeqs2', 'TNT')) |> 
  ggplot(aes(x=Database, y = TP/102*100, fill=Type)) + #fill=factor(Database, levels=c('30%','50%','70%')))) + 
  geom_boxplot(outlier.shape=NA) +
  labs(fill='Type') + 
  ylab('True positives (%)') + 
  geom_point(aes(color=Type), 
           position = position_jitterdodge(jitter.width = 0.2), 
           alpha=0.5,
           show.legend = FALSE)+
  theme_minimal()
true_positives

false_positives/ true_positives + plot_layout(guides = 'collect') & coord_flip()

### Phylogenetic diversity

We can also easily calculate alpha diversity across these tools, as alpha diversity is the number of species. We treat classifiers/Types as sites.

spec_summarised <- twelve_exclusions_split |> 
  group_by(Type, Query, Database, after) |> 
  mutate(Database = case_when ( Database == 'fifty' ~ '50%',
                               Database == 'seventy' ~ '70%',
                               Database == 'thirty' ~ '30%',
                               TRUE ~ Database)) |> 
  filter(!is.na(species)) |> 
  summarise(`Alpha diversity index` = length(unique(species)))
## `summarise()` has grouped output by 'Type', 'Query', 'Database'. You can
## override using the `.groups` argument.
spec_summarised |> 
  ggplot(aes(x = Type, y = `Alpha diversity index`, fill=Type, group = Type )) + 
  geom_boxplot() +
  geom_point(aes(color=Type), 
           position = position_jitterdodge(jitter.width = 0.2), 
           alpha=0.5,
           show.legend = FALSE) + 
  facet_wrap(~Database) + coord_flip() + theme_minimal()

Let’s also do not all of the classifiers

spec_summarised |> 
  filter(Type %in% c('BLAST100', 'Kraken_0.0', 'Kraken_0.05', 'Kraken_0.1','MMSeqs2', 'Qiime2', 'TNT')) |> 
  ggplot(aes(x = Type, y = `Alpha diversity index`, fill=Type, group = Type )) + 
  geom_boxplot() +
  geom_point(aes(color=Type), 
           position = position_jitterdodge(jitter.width = 0.2), 
           alpha=0.5,
           show.legend = FALSE) + 
  facet_wrap(~Database) + coord_flip() + theme_minimal()

a <- spec_summarised |> 
  filter(Type %in% c('BLAST100', 'BLAST97', 'Kraken_0.0','Kraken_0.05', 'Kraken_0.1','MMSeqs2', 'Qiime2', 'TNT')) |> 
  group_by(Database) |> 
  arrange(Database) |> 
  group_map(~aov(`Alpha diversity index` ~ Type, data=.))

names(a) <- spec_summarised |> arrange(Database) |> pull(Database) |> unique() # I don't like this :(
a
## $`30%`
## Call:
##    aov(formula = `Alpha diversity index` ~ Type, data = .)
## 
## Terms:
##                     Type Residuals
## Sum of Squares  4149.933  1070.800
## Deg. of Freedom        5        54
## 
## Residual standard error: 4.453047
## Estimated effects may be unbalanced
## 
## $`50%`
## Call:
##    aov(formula = `Alpha diversity index` ~ Type, data = .)
## 
## Terms:
##                     Type Residuals
## Sum of Squares  3674.483   653.700
## Deg. of Freedom        5        54
## 
## Residual standard error: 3.479304
## Estimated effects may be unbalanced
## 
## $`70%`
## Call:
##    aov(formula = `Alpha diversity index` ~ Type, data = .)
## 
## Terms:
##                     Type Residuals
## Sum of Squares  2567.749   805.975
## Deg. of Freedom        5        52
## 
## Residual standard error: 3.936943
## Estimated effects may be unbalanced
library(agricolae)
groupslist <- list()

for(key in names(a)) {
  print(key)
  groupslist[[key]] <- HSD.test(a[[key]], 'Type')$groups|> 
    as_tibble(rownames = 'Type') |> 
    select(-`Alpha diversity index`)
}
## [1] "30%"
## [1] "50%"
## [1] "70%"
groups_df <- bind_rows(groupslist, .id='Database')
spec_summarised |> 
  filter(Type %in% c('BLAST100', 'BLAST97', 'Kraken_0.0', 'Kraken_0.05', 'Kraken_0.1','MMSeqs2', 'Qiime2', 'TNT')) |> 
  left_join(groups_df, by = c('Database', 'Type')) |> 
  ggplot(aes(x = Type, y = `Alpha diversity index`, fill=Type, group = Type )) + 
  geom_boxplot(outlier.shape=NA) +
  geom_point(aes(color=Type), 
           position = position_jitterdodge(jitter.width = 0.2), 
           alpha=0.5,
           show.legend = FALSE) + 
  facet_wrap(~Database) + 
  geom_text(aes(x = Type, y = max(`Alpha diversity index`) + 2, label = groups),
            #col = 'black',
            size = 4) +
  #coord_flip() + 
  theme_minimal() +
  theme(axis.text.x = element_text( angle = 90, hjust = 1)) +
  guides(fill="none")

16S exclusion databases

sixteen_exclusions <- data |> filter(str_starts(Subject, '16S_v04_final.fasta_Taxonomies.')) |> 
  filter(Query == 'make_12s_16s_simulated_reads_8-Rottnest_runEDNAFLOW_16S_RESULTS_dada2_asv.fa')
table(sixteen_exclusions$Subject)
## 
##    16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_1.fasta 
##                                                                   1269 
##   16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_10.fasta 
##                                                                   1313 
##    16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_2.fasta 
##                                                                   1394 
##    16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_3.fasta 
##                                                                   1290 
##    16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_4.fasta 
##                                                                   1320 
##    16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_5.fasta 
##                                                                   1339 
##    16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_6.fasta 
##                                                                   1348 
##    16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_7.fasta 
##                                                                   1321 
##    16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_8.fasta 
##                                                                   1263 
##    16S_v04_final.fasta_Taxonomies.CountedFams.txt_fifty_subset_9.fasta 
##                                                                   1287 
##  16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_1.fasta 
##                                                                   1411 
## 16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_10.fasta 
##                                                                   1347 
##  16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_2.fasta 
##                                                                   1401 
##  16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_3.fasta 
##                                                                   1334 
##  16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_4.fasta 
##                                                                   1314 
##  16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_5.fasta 
##                                                                   1442 
##  16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_6.fasta 
##                                                                   1380 
##  16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_7.fasta 
##                                                                   1395 
##  16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_8.fasta 
##                                                                   1422 
##  16S_v04_final.fasta_Taxonomies.CountedFams.txt_seventy_subset_9.fasta 
##                                                                   1423 
##   16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_1.fasta 
##                                                                   1263 
##  16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_10.fasta 
##                                                                   1288 
##   16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_2.fasta 
##                                                                   1283 
##   16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_3.fasta 
##                                                                   1267 
##   16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_4.fasta 
##                                                                   1334 
##   16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_5.fasta 
##                                                                   1274 
##   16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_6.fasta 
##                                                                   1234 
##   16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_7.fasta 
##                                                                   1319 
##   16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_8.fasta 
##                                                                   1262 
##   16S_v04_final.fasta_Taxonomies.CountedFams.txt_thirty_subset_9.fasta 
##                                                                   1274
sixteen_exclusions_split <- sixteen_exclusions |> separate(Subject, into = c('before', 'hit'), sep='.txt_') |> 
  # get rid of leftover non-subsetted databases
  filter(!is.na(hit)) |> 
  separate(hit, into=c('Database', 'after'), sep='_subset')
sixteen_exclusions_split_averaged <- sixteen_exclusions_split |> left_join(sixteenS_truth, by = c('OTU' = 'True_OTU')) |>
  separate(True_species, into = c('True_Genus', 'True_Epiteth'), remove = FALSE)|> 
  mutate(species = na_if(species, 'dropped')) |> 
  mutate(genus = na_if(genus, 'dropped')) |> 

  mutate(CorrectSpecies = case_when(!is.na(species) & True_species == species ~ 'Correct species',
                              !is.na(species) & True_species != species ~ 'Incorrect species',
                              !is.na(genus) & !is.na(True_Genus) & True_Genus == genus ~ 'Correct genus',
                              !is.na(genus) & !is.na(True_Genus) & True_Genus != genus ~ 'Incorrect genus',
                              !is.na(family) & True_family == family ~ 'Correct family',
                              !is.na(family) & True_family != family ~ 'Incorrect family',
                              TRUE ~ NA)) |> 
  group_by(Type, Database, after) |>
  summarise(TP = sum(str_detect(CorrectSpecies, pattern='Correct species'), na.rm=TRUE),
            FP = sum(str_detect(CorrectSpecies, pattern = 'Incorrect species'), na.rm=TRUE),
            TN = sum(str_detect(replace_na(CorrectSpecies,'NA'), pattern='NA') & is.na(True_species), na.rm=TRUE),
            FN = sum(is.na(species) & !is.na(True_species))) |> 
  mutate(sums = TP + FP + TN + FN) |> 
  mutate(missing = 102 - sums) |> 
  mutate(FN = FN + missing) |> 
  mutate(sums = TP + FP + TN + FN) |> 
  select(-c(missing, sums)) |> 
  group_by(Type, Database) |> 
  summarise(mean_TP = mean(TP),
            mean_FP = mean(FP),
            mean_TN = mean(TN),
            mean_FN = mean(FN)) |> 
  rowwise() |> 
  mutate(recall = recall(mean_TP, mean_FN), 
         precision = precision(mean_TP, mean_FP),
         f1 = f1(precision, recall),
         f0.5 = f0.5(precision, recall),
         accuracy = accuracy(mean_TP, mean_FP, mean_FN, mean_TN)) 
## `summarise()` has grouped output by 'Type', 'Database'. You can override using
## the `.groups` argument.
## `summarise()` has grouped output by 'Type'. You can override using the
## `.groups` argument.
sixteen_exclusions_split_averaged <- sixteen_exclusions_split_averaged |> 
  mutate(Database = case_when ( Database == 'fifty' ~ '50%',
                               Database == 'seventy' ~ '70%',
                               Database == 'thirty' ~ '30%',
                               TRUE ~ Database))
f1_pl <- sixteen_exclusions_split_averaged |> 
  ggplot(aes(x = Type, y = f1, group = Database, color = Database, fill = Database)) + 
  geom_point() +
  geom_line() +
  theme_minimal()
f0.5_pl <- sixteen_exclusions_split_averaged |> 
  ggplot(aes(x = Type, y = f0.5, group = Database, color = Database, fill = Database)) + 
  geom_point() +
  geom_line() +
  theme_minimal()
precision_pl <- sixteen_exclusions_split_averaged |> 
  ggplot(aes(x = Type, y = precision, group = Database, color = Database, fill = Database)) + 
  geom_point() +
  geom_line() +
  theme_minimal()
recall_pl <- sixteen_exclusions_split_averaged |> 
  ggplot(aes(x = Type, y = recall, group = Database, color = Database, fill = Database)) + 
  geom_point() +
  geom_line() +
  theme_minimal()
(f1_pl / f0.5_pl / precision_pl / recall_pl) +  plot_layout(guides = 'collect')

Lets zero in on the precision and make boxplots with jitter dots

un_summarised_sixteen <- sixteen_exclusions_split |> left_join(sixteenS_truth, by = c('OTU' = 'True_OTU')) |>
  separate(True_species, into = c('True_Genus', 'True_Epiteth'), remove = FALSE)|> 
  mutate(species = na_if(species, 'dropped')) |> 
  mutate(genus = na_if(genus, 'dropped')) |> 

  mutate(CorrectSpecies = case_when(!is.na(species) & True_species == species ~ 'Correct species',
                              !is.na(species) & True_species != species ~ 'Incorrect species',
                              !is.na(genus) & !is.na(True_Genus) & True_Genus == genus ~ 'Correct genus',
                              !is.na(genus) & !is.na(True_Genus) & True_Genus != genus ~ 'Incorrect genus',
                              !is.na(family) & True_family == family ~ 'Correct family',
                              !is.na(family) & True_family != family ~ 'Incorrect family',
                              TRUE ~ NA)) |> 
  group_by(Type, Database, after) |>
  summarise(TP = sum(str_detect(CorrectSpecies, pattern='Correct species'), na.rm=TRUE),
            FP = sum(str_detect(CorrectSpecies, pattern = 'Incorrect species'), na.rm=TRUE),
            TN = sum(str_detect(replace_na(CorrectSpecies,'NA'), pattern='NA') & is.na(True_species), na.rm=TRUE),
            FN = sum(is.na(species) & !is.na(True_species))) |> 
  mutate(sums = TP + FP + TN + FN) |> 
  mutate(missing = 112 - sums) |> 
  mutate(FN = FN + missing) |> 
  mutate(sums = TP + FP + TN + FN) |> 
  select(-c(missing, sums)) |> 
  rowwise() |> 
  mutate(recall = recall(TP, FN), 
         precision = precision(TP, FP),
         f1 = f1(precision, recall),
         f0.5 = f0.5(precision, recall),
         accuracy = accuracy(TP, FP, FN, TN)) |> 
  mutate(Database = case_when ( Database == 'fifty' ~ '50%',
                               Database == 'seventy' ~ '70%',
                               Database == 'thirty' ~ '30%',
                               TRUE ~ Database))
## `summarise()` has grouped output by 'Type', 'Database'. You can override using
## the `.groups` argument.
un_summarised_sixteen |> group_by(Type, Database) |> mutate(best = max(mean(precision, na.rm = TRUE))) |>
  ggplot(aes(x = fct_reorder(Type, best), group = interaction(Type, Database),  color=Database, y = precision)) +
  geom_boxplot(outlier.shape = NA) +
  coord_flip() +
  theme_minimal() +
  xlab('Type') +
  ylab('Precision') +
  geom_point(position = position_jitterdodge(), alpha=0.5)

un_summarised_sixteen  |> group_by(Type, Database) |> mutate(best = max(mean(f0.5, na.rm=TRUE))) |>
  ggplot(aes(x = fct_reorder(Type, best), group = interaction(Type, Database), color=Database, y = f0.5)) +
  geom_boxplot(outlier.shape = NA) + 
  coord_flip() + 
  theme_minimal() + 
  xlab('Type') +
  ylab('f0.5') +
  ylim(c(0, 1)) +
  geom_point(position = position_jitterdodge(), alpha=0.5)

un_summarised_sixteen  |> group_by(Type, Database) |> mutate(best = max(mean(recall))) |>
  ggplot(aes(x = fct_reorder(Type, best), group = interaction(Type, Database), color=Database, y = recall)) +
  geom_boxplot(outlier.shape = NA) + 
  coord_flip() + 
  theme_minimal() + 
  xlab('Type') +
  ylab('f0.5') +
  ylim(c(0, 1)) +
  geom_point(position = position_jitterdodge(), alpha=0.5)

un_summarised_sixteen  |> 
  filter(Type %in% c('BLAST100', 'Kraken_0.0',  'Kraken_0.05', 'Qiime2', 'TNT')) |> 
  ggplot(aes(x=Database, y = precision, fill=Type)) + #fill=factor(Database, levels=c('30%','50%','70%')))) + 
  geom_boxplot() +
  labs(fill='Type') + 
  ylab('Precision') + 
  theme_minimal()

false_positives <- un_summarised_sixteen  |> 
  filter(Type %in% c('BLAST100', 'Kraken_0.0',  'Kraken_0.05', 'Kraken_0.1', 'MMSeqs2', 'TNT')) |> 
  ggplot(aes(x=Database, y = FP/112*100, fill=Type)) + #fill=factor(Database, levels=c('30%','50%','70%')))) + 
  geom_boxplot(outlier.shape=NA) +
  labs(fill='Type') + 
  ylab('False positives (%)') + 
  geom_point(aes(color=Type), 
             position = position_jitterdodge(jitter.width = 0.2), 
             alpha=0.5,
             show.legend = FALSE)+
  theme_minimal()
false_positives

true_positives <- un_summarised_sixteen  |> 
  filter(Type %in% c('BLAST100', 'Kraken_0.0',  'Kraken_0.05', 'Kraken_0.1', 'MMSeqs2', 'TNT')) |> 
  ggplot(aes(x=Database, y = TP/112*100, fill=Type)) + #fill=factor(Database, levels=c('30%','50%','70%')))) + 
  geom_boxplot(outlier.shape=NA) +
  labs(fill='Type') + 
  ylab('True positives (%)') + 
  geom_point(aes(color=Type), 
           position = position_jitterdodge(jitter.width = 0.2), 
           alpha=0.5,
           show.legend = FALSE)+
  theme_minimal()
true_positives

false_positives/ true_positives + plot_layout(guides = 'collect') & coord_flip()

### Phylogenetic diversity

We can also easily calculate alpha diversity across these tools, as alpha diversity is the number of species. We treat classifiers/Types as sites.

spec_summarised <- sixteen_exclusions_split |> 
  group_by(Type, Query, Database, after) |> 
  mutate(Database = case_when ( Database == 'fifty' ~ '50%',
                               Database == 'seventy' ~ '70%',
                               Database == 'thirty' ~ '30%',
                               TRUE ~ Database)) |> 
  filter(!is.na(species)) |> 
  summarise(`Alpha diversity index` = length(unique(species)))
## `summarise()` has grouped output by 'Type', 'Query', 'Database'. You can
## override using the `.groups` argument.
spec_summarised |> 
  ggplot(aes(x = Type, y = `Alpha diversity index`, fill=Type, group = Type )) + 
  geom_boxplot() +
  geom_point(aes(color=Type), 
           position = position_jitterdodge(jitter.width = 0.2), 
           alpha=0.5,
           show.legend = FALSE) + 
  facet_wrap(~Database) + coord_flip() + theme_minimal()

Let’s also do not all of the classifiers

spec_summarised |> 
  filter(Type %in% c('BLAST100', 'Kraken_0.0', 'Kraken_0.05',  'Kraken_0.1','MMSeqs2', 'Qiime2', 'TNT')) |> 
  ggplot(aes(x = Type, y = `Alpha diversity index`, fill=Type, group = Type )) + 
  geom_boxplot() +
  geom_point(aes(color=Type), 
           position = position_jitterdodge(jitter.width = 0.2), 
           alpha=0.5,
           show.legend = FALSE) + 
  facet_wrap(~Database) + coord_flip() + theme_minimal()

a <- spec_summarised |> 
  filter(Type %in% c('BLAST100', 'BLAST97', 'Kraken_0.0',  'Kraken_0.05', 'Kraken_0.1','MMSeqs2', 'Qiime2', 'TNT')) |> 
  group_by(Database) |> 
  arrange(Database) |> 
  group_map(~aov(`Alpha diversity index` ~ Type, data=.))

names(a) <- spec_summarised |> arrange(Database) |> pull(Database) |> unique() # I don't like this :(
a
## $`30%`
## Call:
##    aov(formula = `Alpha diversity index` ~ Type, data = .)
## 
## Terms:
##                     Type Residuals
## Sum of Squares  8446.646  1729.975
## Deg. of Freedom        5        52
## 
## Residual standard error: 5.767907
## Estimated effects may be unbalanced
## 
## $`50%`
## Call:
##    aov(formula = `Alpha diversity index` ~ Type, data = .)
## 
## Terms:
##                     Type Residuals
## Sum of Squares  6966.745  3111.000
## Deg. of Freedom        5        49
## 
## Residual standard error: 7.968048
## Estimated effects may be unbalanced
## 
## $`70%`
## Call:
##    aov(formula = `Alpha diversity index` ~ Type, data = .)
## 
## Terms:
##                     Type Residuals
## Sum of Squares  4175.647   788.914
## Deg. of Freedom        5        51
## 
## Residual standard error: 3.933053
## Estimated effects may be unbalanced
library(agricolae)
groupslist <- list()

for(key in names(a)) {
  print(key)
  groupslist[[key]] <- HSD.test(a[[key]], 'Type')$groups|> 
    as_tibble(rownames = 'Type') |> 
    select(-`Alpha diversity index`)
}
## [1] "30%"
## [1] "50%"
## [1] "70%"
groups_df <- bind_rows(groupslist, .id='Database')
spec_summarised |> 
  filter(Type %in% c('BLAST100', 'BLAST97', 'Kraken_0.0', 'Kraken_0.05',  'Kraken_0.1','MMSeqs2', 'Qiime2', 'TNT')) |> 
  left_join(groups_df, by = c('Database', 'Type')) |> 
  ggplot(aes(x = Type, y = `Alpha diversity index`, fill=Type, group = Type )) + 
  geom_boxplot(outlier.shape=NA) +
  geom_point(aes(color=Type), 
           position = position_jitterdodge(jitter.width = 0.2), 
           alpha=0.5,
           show.legend = FALSE) + 
  facet_wrap(~Database) + 
  geom_text(aes(x = Type, y = max(`Alpha diversity index`) + 2, label = groups),
            #col = 'black',
            size = 4) +
  #coord_flip() + 
  theme_minimal() +
  theme(axis.text.x = element_text( angle = 90, hjust = 1)) +
  guides(fill="none")